Hansen bioprotect wines with viniflora

1. Bioprotect musts, wines and brands with NSAC yeasts
ACE – Arnes, 2015
Bioprotect musts, wines and brands with NSAC yeasts
ACE – Arnes, 2015
• --

2. ct ’OK’
Bio-protective
effects
against contaminants
Natural way to increase
fermentation flavors
intensity, longevity and complexity
Full use of
grape juice and wine
bio-diversity
No production
of biogenic amines
Better management
of grape juice initial content:
flavor precursors/org. acids/…
Excellent
return on investment
Less downgrades
during production
=
preserve initial potential
and personality
Outline: feedback from winemakers using NSAC yeasts
Why is it interesting to use NSAC yeasts? 6 years global survey
Outline: feedback from winemakers using NSAC yeasts
Why is it interesting to use NSAC yeasts? 6 years global survey

3. NSAC under scrutiny: the starting point…NSAC under scrutiny: the starting point…
Population dynamics of non-Saccharomyces yeast
Goddard MR. 2008. Ecology 89: 2077-2082
NSAC
proliferate at
early stages
Saccharomyces takes
over, after starting from
a low conc.

4. ct ’OK’
MFC* sequence in winemakingMFC* sequence in winemaking
Population
Time
in days then weeks
Non-Saccharomyces
spp.
Saccharomyces
spp. Oenococcus oeni /
Lactobacillus ssp.
Alcoholic
fermentation
Malolactic
fermentation
* MFC: Microbial Food Cultures

5. Metabolites produced by non-Saccharomyces yeasts
Non-Saccharomyces wine yeasts are mainly studied for their impact
on flavors, glycerol and organic acids production… and their potential
ability to convert sugars into these metabolites instead of ethanol
Number of peer reviewed publications and reviews
per wine non-Sacch. positive to neutral species and main metabolites studied
period 2005-2012
0
5
10
15
20
25
30
Torulaspora
Kluyveromyces
Pichia
Metschnikowia
Schizosaccharomyces
Hanseniaspora

6. 6
NSAC yeasts: organoleptic impact is the primary aim
wine characteristics impacted per type
1 species among wine NSAC yeast
has shown the highest
potential: Pichia kluyveri
1 species among wine NSAC yeast
has shown the highest potential:
Lachancea* thermotolerans
* : following OIV guidelines regarding wine yeast selection
* *: formerly Kluyveromyces thermotolerans
1 species among wine NSAC yeast
has shown the highest
potential: Torulaspora delbrueckii
Flavors
Precursors conversion
Metabolism differences
Acid balance
Organic acids
production/reduction
Mouth-feel
Polysaccharides
Mannoproteins
Concerto™ Prelude™FrootZen™

7. NSAC yeasts: nowadays protection against contaminants is
becoming the primary objective of their application
Phase 1 Phase 2
2003
Pure strains
Bio-protection
New applications
Phase 3
2009
Phase 4
2012
BlendsFirst attempts
- Torulaspora
delbrueckii
- Schizosaccharomyces
pombe
- …
- Saccharomyces
cerevisiae,
mixed with:
- Torulaspora
delbrueckii
and/or
- Kluyveromyces
thermotolerans
MELODY™ blend
from Chr. Hansen
- Torulaspora
delbrueckii
- Lachancea/Kluyveromyces
thermotolerans
- Pichia kluyveri
- …
Mimic wild ferment
Add extra complexity
Initial interest for
exotic yeast species
potential
Get ‘wild ferment’
benefits without
the risks
Preserve, protect, convert
precursors contained in must
and avoid contaminants
PRELUDE™ first pure Td
product launched in August
2009 by Chr. Hansen
Ex.
of
species
Main
use
Ex. of
products
FrootZen™ first pure Pk
product by Chr. Hansen
Timeline

8. NSAC yeasts: bio-protection mainly comes from competitive
exclusion, 7 different applications have been developed
* MFC: Microbial Food Cultures
NSAC yeasts new applications in winemaking:
COMPETITIVE EXCLUSION
NSAC inoculation: high concentration per ml of viable populations
from species that are well adapted to must conditions (high sugar
content for instance) vs. Saccharomyces spp.
=> fast development that will limit availability of nutrients for
molds/other yeasts/ bacteria and create the few % ethanol to
protect the must
Advanced winemakers are now using
this competitive exclusion feature
to bio-protect red, rosé or white musts
4 main final objectives identified
No/less sulfites in winemaking
<10 ppm sulfites in bottles
Avoid Biogenic amines
(histamine/tyramine/…)
Avoid phenolic compounds/
volatile acidity
Limit oxygen impact
Optimize flavor precursors conversion
(pool of enzymes, early timing, …)
Bring complexity (flavors/org.
acids/polysaccharides/ …)
When used earlier enough in the process, spread on grapes right
after crush for instance, high populations of some NSAC yeasts bring
protection against contaminants by competitive exclusion
(competing to access the same limited resources than molds, wild
yeasts, some bacteria…)
Reduce downgrades
Preserve wines´ personality

9. 14 April 2015 Australia & NZ, July 20129
Viniflora® NSAC yeasts: impact on mouthfeel
Flavours
Precursors conversion
Metabolism differences
Acid balance
Organic acids
production/reduction
Mouth-feel
Polysaccharides
Concerto™ Prelude™FrootZen™

10. ct ’OK’
Viniflora® PRELUDE™Viniflora® PRELUDE™

11. 14 April 2015 Australia & NZ, July 201211
Torulaspora delbrueckii - effect on palate-weight:
high production of polysaccharides
Sc alone Sc + Torulaspora delbrueckii
Source: Comitini, F., et al., Selected non-Saccharomyces wine yeasts in controlled multistarter f..., Food Microbiology (2011)

12. 12
Viniflora® PRELUDE™ usage in cold soak: to protect the must
by competitive exclusion
Average population of cells (in CFU/ml) for Prelude™ (orange),
Frootzen™ (blue) and the blend of NSAC and SAC Melody (green) on YPG media
at different temperatures
Implantation and growth in musts at low temperature
of different NSAC products

13. Viniflora® Prelude™ - Torulaspora delbrueckii
example of volatile acidity reduction & fatty acids reduction
13
Acetic acid
g/l
Glycerol
g/l
Esters
mg/l
Fatty acids (C6,8,10,12)
mg/l
S. cerevisiae (EC1118) 0.21 6.7 6.3 14.5
T. delbrueckii (Prelude™)
+
S. cerevisiae (EC1118)
0.06 6.9 7.3 3.5
p.t. ~ 20 mg/L
The primary reason for these decrease in
volatile acidity and fatty acids is the high
tolerance of Td yeasts for high sugar substrates,
they are less stressed by their environment than
Saccharomyces spp.

14. 14
0
10
20
30
40
50
60
70
Merit + CiNe Prelude + Merit + CiNE FrootZen + Merit + CiNe
Percent[%]malicacidconcumed
Day 2
Day 4
Viniflora® PRELUDE™ : effect of treating the must with T.
delbrueckii on MLF management
Malic acid degradation in %
measured 2 and 4 days after inoculation with Viniflora® CiNe™
Merit: pure S. cerevisiae, control
Prelude: pure T. delbrueckii
FrootZen: pure P. kluyveri

15. 15
Customers in 15 countries
Primarily in Europe, South Africa, New Zealand, Chile, USA
Dosages vary from 30 to 60 g/hl; 2 days to 8 days ferment.
Red wines or white wines matured with oak chips or in barrels
Red wines grapes picked early (to limit ethanol final concentration)
Cold soak/pre fermentation maceration (Pinot Noir for instance)
Prelude™ (Td CH111) applications

16. 14 April 2015 Australia & NZ, July 201216
Viniflora® NSAC yeasts: impact on flavors
Concerto™ Prelude™FrootZen™
Flavors
Precursors conversion
Metabolism differences
Acid balance
Organic acids
production/reduction
Mouth-feel
Polysaccharides

17. ct ’OK’
Viniflora® FROOTZEN™Viniflora® FROOTZEN™

18. FrootZen™: the magic of direct inoculation

19. Sequential inoculation: FrootZen™ followed by S. cerevisiae
Time
Must
Specific gravity
° Brix
Direct inoculation
with FrootZen™
Inoculation
with usual standard
S. cerevisiae yeast
1100
24°
B
990
1070
17°
B
Wine
Wine ready for
malolactic fermentation
with Viniflora®
cfu/ml
of wine
1E+04
1E+05
0
1E+06
1030
8°B
30 points
7 °°°°B
STEP 1
STEP 2
5E+05 cfu/ml at inoculation

20. Additional effects of FrootZen™ protecting the musts
faster consumption of oxygen than S. cerevisiae
Additional effects of FrootZen™ protecting the musts
faster consumption of oxygen than S. cerevisiae

21. Sugar consumption
0,0
20,0
40,0
60,0
80,0
100,0
120,0
140,0
160,0
180,0
200,0
0 1 2 3 4 5 6 7
Day
FrucandGlucg/L
Sc
Td
Pk
Viniflora® FrootZen™ : fermentation kinetic
key differences compare to Sc and Td
Viniflora® FrootZen™ : fermentation kinetic
key differences compare to Sc and Td

22. Ethanol production
0,0
5,0
10,0
15,0
0 1 2 3 4 5 6 7
Day
Ethanolvol%
Sc
Td
Pk
Viniflora® FrootZen™ : fermentation kinetic
Pichia kluyveri is not a strong fermenter, ethanol production is limited
Viniflora® FrootZen™ : fermentation kinetic
Pichia kluyveri is not a strong fermenter, ethanol production is limited

23. Total acid
2,0
2,5
3,0
3,5
4,0
0 1 2 3 4 5 6 7
Day
g/L
Sc
Td
Pk
Viniflora® FrootZen™ : fermentation kinetic
Pichia kluyveri produces organic acids… and flavors!
Viniflora® FrootZen™ : fermentation kinetic
Pichia kluyveri produces organic acids… and flavors!

24. FrootZen™ impact on consumer panels:
value creation
Measure with a panels:
professionals and/or consumers
Assess through these information:
How the wine made with FrootZen™ is rated
and the value added by the NSAC yeast
compare to the control
Demonstrate the added value created by the
NSAC yeast
Generalize its use in wineries to differentiate
wines and improve their value.
In this example: > 40% price difference
between real price and jury estimated price
24
HOW TO EXPLAIN THIS SYSTEMATIC
CREATION OF VALUE?

25. Flavors frequence mentioned by external trained/non professional jury
Loire Valley wine (Sancerre - Sauvignon blanc)
Data: In senso veritas, Feb. 2013
Tropical fruit notes Floral notes
Sensory profile improvement with FrootZen™
white wine example (Sauvignon blanc)
Sensory profile improvement with FrootZen™
white wine example (Sauvignon blanc)

26. Sensory profile improvement with FrootZen™
red wine example (Pinot Noir)
Sensory profile improvement with FrootZen™
red wine example (Pinot Noir)

27. 27
Customers in 20 countries
Primarily in Europe, South Africa, New Zealand, USA
2 to 4 days in fermentation.
Application in red/white/rosé wines
Intense, elegant wines, very early protection against contaminants
FrootZen™ (Pk) applications

28. 14 April 2015 Australia & NZ, July 201228
Viniflora® NSAC yeasts: impact on organic acids balance
Flavours
Precursors conversion
Metabolism differences
Acid balance
Organic acids
production/reduction
Mouth-feel
Polysaccharides
Concerto™ Prelude™FrootZen™

29. ct ’OK’
Viniflora® CONCERTO™Viniflora® CONCERTO™

30. Concerto™ - Kluyveromyces thermotolerans – strain CH456
lactic acid production -natural acidification- ideal for
warm climates red or rosé wines
Sc only
control
Kluyveromyces* thermotolerans
+ Sc
Source: Comitini, F., et al., Selected non-Saccharomyces wine yeasts in controlled multistarter f..., Food Microbiology (2011)
* : Kluyveromyces thermotolerans is also named Lachancea thermotolerans (L. thermotolerans)

31. Australia & NZ, July 201231
Concerto™ vs. FrootZen™ vs. Prelude™
Trials carried out in 2011 to assess the differences imparted by 3 pure NSAC
Prelude™, FrootZen™ and Concerto™
Three wines: Red, Rosé and White
Bodegas Purisima Concepción, Ribera del Jucar, Spain
Where?
How? SEQUENTIAL INOCULATION: NSAC first followed by SAC
NSAC inoculated first
Dosage 20g/hl for ADY
today we recommend 25g/hl for
ADY
Dosage 10g/hl for FROOTZEN™
CIU for winemaker: 2cEuro/L
ROI monitored,
average measured: 2E/L
SAC inoculated
2 different yeasts for white
and rosé
Dosage 15g/hl
SAC inoculated after a loss of
25-30 points in spec. gravity
36 to 48h most time.

32. 14 April 2015 Australia & NZ, July 201232
The Red: Cabernet Sauvignon
0
1
2
3
4
5
6
7
Acidity
Fruitiness
Body
Bitterness
Cherry
Black plum
Strawberry
Blackberry
Honey
Green Beans
Cabernet Sauvignon
FrootZen
Prelude
Concerto
Winemaking:
Sacc: S.G. 101
AF temp: 25°C
Maceration: 6 days
MLF: Yes
Analysis:
RS: 3.0 g/l
Alc: 13.8%
pH 3.6
TA = 4.8g/l
(as tartaric)

33. 14 April 2015 Australia & NZ, July 201233
The Rosé: Bobal
Rosé from Bobal
0
1
2
3
4
5
6
7
Acidity
Fruitiness
Body
Bitterness
CherryBlack plum
Strawberry
Blackberry
Green Beans
FrootZen
Prelude
Concerto
Winemaking:
Sacc: S.G. 101
AF temp: 16°C
Maceration: 4 hours
MLF: No
Analysis:
RS: 3.5 g/l
Alc: 14.0%
pH 3.5
TA = 4.8g/l
(as tartaric)

34. 34
Customers in 10 countries
Primarily in Europe, South Africa, Australia, USA
Dosages vary from 30 to 60 g/hl; 2 days to 10 days ferment.
Red/white/rosé wines from warm climate areas
Cleaner wines, more complexity, far less contamination issues
Concerto™ (Kt CH456) applications

35. Conclusions
No/less sulfites
in winemaking
<10 ppm sulfites in bottles
Avoid Biogenic amines
(histamine/tyramine/…)
Avoid phenolic compounds/
volatile acidity
Limit oxygen impact
Optimize flavor precursors
conversion (pool of enzymes,
early timing, …)
Bring complexity (flavors/org.
acids/polysaccharides/ …)
Reduce downgrades
Preserve wines´
personality
Flavours
Precursors conversion
Metabolism diff.
Acid balance
Organic acids
production/reduction
Mouth-feel
Polysaccharides
Concerto™ Prelude™FrootZen™
++ + +
++ ++
++ ++ ++
Historical
primary aim
during strain selection

36. 36

37. --
Thank you for your attention!
more information are available at
www.chr-hansen.com/wine
Thank you for your attention!
more information are available at
www.chr-hansen.com/wine